Name: Anti-AMF antibody [EPR11663(B)]
Brand: RabMAb
SKU: ab167394
Rating: 4 (1 reviews)

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Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR11663(B)] to AMF
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
Knockout validated
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free PE

Product name

Anti-AMF antibody [EPR11663(B)]
See all AMF primary antibodies
Description

Rabbit monoclonal [EPR11663(B)] to AMF
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide corresponding to Human AMF.
Database link: P06744
Positive control
- WB: HEK-293T, HepG2, HeLa, HT29, and A549 cell lysates. IHC-P: Human brain, pancreas, skeletal muscle, ovarian carcinoma, prostatic hyperplasia and thyroid carcinoma tissues. ICC/IF: MCF7 and HepG2 cells. Flow Cyt (intra): A549 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR11663(B)
Isotype

IgG
Research areas

Alternative Versions
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human GPI (AMF) knockout HEK-293T cell line (ab266834)
KO cell lysates
- Human GPI (AMF) knockout HEK-293T cell lysate (ab257458)
KO cell pellets
- Human GPI (AMF) knockout HEK-293T cell pellet (ab278990)
Positive Controls
- HeLa whole cell lysate (ab29545)
- A549 whole cell lysate (ab7910)
Recombinant Protein
- Recombinant Human AMF protein (ab87625)
Related Products
- Recombinant Human AMF protein (ab87625)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab167394 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB	(1)	1/1000 - 1/10000. Predicted molecular weight: 63 kDa.
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF		1/100 - 1/500.

Notes
Flow Cyt (Intra) 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/10000. Predicted molecular weight: 63 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/500.

Application notes

Is unsuitable for IP.

Function

Besides it's role as a glycolytic enzyme, mammalian GPI can function as a tumor-secreted cytokine and an angiogenic factor (AMF) that stimulates endothelial cell motility. GPI is also a neurotrophic factor (Neuroleukin) for spinal and sensory neurons.
Pathway

Carbohydrate degradation; glycolysis; D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose: step 2/4.
Involvement in disease

Defects in GPI are the cause of hemolytic anemia non-spherocytic due to glucose phosphate isomerase deficiency (HA-GPID) [MIM:613470]. It is a form of anemia in which there is no abnormal hemoglobin or spherocytosis. It is caused by glucose phosphate isomerase deficiency. Severe GPI deficiency can be associated with hydrops fetalis, immediate neonatal death and neurological impairment.
Sequence similarities

Belongs to the GPI family.
Post-translational
modifications

Phosphorylation at Ser-185 by CK2 has been shown to decrease enzymatic activity and may contribute to secretion by a non-classical secretory pathway.
ISGylated.
Cellular localization

Cytoplasm. Secreted.
Target information above from: UniProt accession P06744 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2821 Human
- Omim: 172400 Human
- SwissProt: P06744 Human
- Unigene: 466471 Human
Alternative names
- AMF antibody
- Aurocrine motility factor antibody
- Autocrine motility factor antibody
- DKFZp686C13233 antibody
- EC 5.3.1.9 antibody
- G6PI_HUMAN antibody
- Glucose phosphate isomerase antibody
- Glucose-6-phosphate isomerase antibody
- GNPI antibody
- GPI antibody
- Gpi1 antibody
- Hexose monophosphate isomerase antibody
- Hexosephosphate isomerase antibody
- Neuroleukin antibody
- NLK antibody
- Oxoisomerase antibody
- PGI antibody
- PHI antibody
- Phosphoglucose isomerase antibody
- Phosphohexomutase antibody
- Phosphohexose isomerase antibody
- Phosphosaccharomutase antibody
- SA 36 antibody
- SA-36 antibody
- SA36 antibody
- Sperm antigen 36 antibody
see all

Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)

All lanes : Anti-AMF antibody [EPR11663(B)] (ab167394) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : GPI knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 63 kDa
Observed band size: 63 kDa

Lanes 1-4: Merged signal (red and green). Green - ab167394 observed at 63 kDa. Red - loading control ab8245 observed at 36 kDa.

ab167394 Anti-AMF antibody [EPR11663(B)] was shown to specifically react with AMF in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266834 (knockout cell lysate ab257458) was used. Wild-type and AMF knockout samples were subjected to SDS-PAGE. ab167394 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [EPR11663(B)] (ab167394)

Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling AFM with purified ab167394 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

Control: PBS only
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

ab167394 showing +ve staining in Human normal brain.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-AMF antibody [EPR11663(B)] (ab167394)

Intracellular flow cytometric analysis of permeabilized A549 cells, labeling AFM with ab167394 at 1/10 dilution.
Western blot - Anti-AMF antibody [EPR11663(B)] (ab167394)

All lanes : Anti-AMF antibody [EPR11663(B)] (ab167394) at 1/1000 dilution

Lane 1 : HepG2 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HT29 cell lysate
Lane 4 : A549 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 63 kDa
Immunocytochemistry/ Immunofluorescence - Anti-AMF antibody [EPR11663(B)] (ab167394)

Immunofluorescent analysis of HepG2 cells labeling AFM with ab167394 at 1/100 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

ab167394 showing +ve staining in Human normal pancreas.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

ab167394 showing +ve staining in Human ovarian carcinoma.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

ab167394 showing +ve staining in Human skeletal muscle.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue, labeling AFM with ab167394 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMF antibody [EPR11663(B)] (ab167394)

Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling AFM with ab167394 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Anti-AMF antibody [EPR11663(B)] (ab167394)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab167394? Please let us know so that we can cite the reference in this datasheet.

ab167394 has been referenced in 1 publication.

He Q et al. Mesenchymal stem cell-derived exosomes exert ameliorative effects in type 2 diabetes by improving hepatic glucose and lipid metabolism via enhancing autophagy. Stem Cell Res Ther 11:223 (2020). PubMed: 32513303

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Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa)

Gel Running Conditions

Reduced Denaturing

Loading amount

83000 cells

Specification

HeLa

Blocking step

Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Nov 12 2021

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

KO cell lines

KO cell lysates

KO cell pellets

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Pathway

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (1)

Customer reviews and Q&As

Western blot abreview for Anti-AMF antibody [EPR11663(B)]

Post-translational
modifications